【求助】能否用ELISA法检测基因突变吗？
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请问能用ELISA（酶连接免疫吸附）法检测基因突变吗？感激不尽！
ELISA-based Ki-ras gene mutation analyses in pancreatic and cholangiocarcinoma cells and tissues.
Abstract
Detection of Ki-ras mutations is one possible modality for diagnosis of human neoplasms, particularly of the gastrointestinal tract. Little is known about the protein expression levels of ras p21. Here we show a semi-quantitative analysis of mutated ras p21 in protein extracts from pancreatic and cholangiocarcinoma cell lines and from tumor tissues. Comparison with DNA sequencing data confirmed the specificity of the ELISA-mediated mutation analysis. Epithelial cell content from stroma rich tissues was estimated by measuring the CK 8, 18 and 19 concentration in protein extracts from tissue samples comparing this with the average keratin content in extracts from 21 GI-carcinoma cell lines. The combination of both tests allowed the analysis of the ras p21 content of tissues with as little as 5% tumor cell content.
Abstract
Detection of Ki-ras mutations is one possible modality for diagnosis of human neoplasms, particularly of the gastrointestinal tract. Little is known about the protein expression levels of ras p21. Here we show a semi-quantitative analysis of mutated ras p21 in protein extracts from pancreatic and cholangiocarcinoma cell lines and from tumor tissues. Comparison with DNA sequencing data confirmed the specificity of the ELISA-mediated mutation analysis. Epithelial cell content from stroma rich tissues was estimated by measuring the CK 8, 18 and 19 concentration in protein extracts from tissue samples comparing this with the average keratin content in extracts from 21 GI-carcinoma cell lines. The combination of both tests allowed the analysis of the ras p21 content of tissues with as little as 5% tumor cell content.
呵呵，这个ELISA检测基因突变比较意思。刚刚看了几篇文章，跟楼主分享一下。
文章：Detection of four b-thalassemia point mutations in Iranians using a PCR-ELISA genotyping system.
检测图中四个位置的基因突变。这个检测方法的四要素：a，DIG labed PCR products b，5’biotinylated oligos (probe) c，streptavidin-coated microtiter d，HRP conjugate anti-digoxigenin antibody 。 检测方法的本质是基于：a，allele-specific oligonucleotide hybridization b，biotin-streptavidin system c，Enzyme-linked immunosorbent assay
Fig 1 - Detection of four b-thalassemia point mutations in Iranians using a PCR-ELISA genotyping system.
文章Molecular Detection of Galactosemia Mutations by PCR-ELISA 有step by step的protocol。
Fig 2 - Molecular Detection of Galactosemia Mutations by PCR-ELISA
PCR-ELISA的原理用于分子诊断方面：TrimGen公司的KRAS Mutation Detection Kit在96孔板里一次能实现7/8个KRAS突变位点的检测。
另外基于相同原理的技术还有PPT-ELISA。这是用来筛截短突变的，相关文章见：1，Rapid screen for truncating ATM mutations by PTT-ELISA . 2，An ELISA-based high throughput protein truncation test for inherited breast cancer .
Fig 3 - Rapid screen for truncating ATM mutations by PTT-ELISA
Fig 4 - An ELISA-based high throughput protein truncation test for inherited breast cancer
文章：Detection of four b-thalassemia point mutations in Iranians using a PCR-ELISA genotyping system.
检测图中四个位置的基因突变。这个检测方法的四要素：a，DIG labed PCR products b，5’biotinylated oligos (probe) c，streptavidin-coated microtiter d，HRP conjugate anti-digoxigenin antibody 。 检测方法的本质是基于：a，allele-specific oligonucleotide hybridization b，biotin-streptavidin system c，Enzyme-linked immunosorbent assay
Fig 1 - Detection of four b-thalassemia point mutations in Iranians using a PCR-ELISA genotyping system.
文章Molecular Detection of Galactosemia Mutations by PCR-ELISA 有step by step的protocol。
Fig 2 - Molecular Detection of Galactosemia Mutations by PCR-ELISA
PCR-ELISA的原理用于分子诊断方面：TrimGen公司的KRAS Mutation Detection Kit在96孔板里一次能实现7/8个KRAS突变位点的检测。
另外基于相同原理的技术还有PPT-ELISA。这是用来筛截短突变的，相关文章见：1，Rapid screen for truncating ATM mutations by PTT-ELISA . 2，An ELISA-based high throughput protein truncation test for inherited breast cancer .
Fig 3 - Rapid screen for truncating ATM mutations by PTT-ELISA
Fig 4 - An ELISA-based high throughput protein truncation test for inherited breast cancer
Fig2 - Molecular Detection of Galactosemia Mutations by PCR-ELISA
Fig3 - Rapid screen for truncating ATM mutations by PTT-ELISA
Fig4 - An ELISA-based high throughput protein truncation test for inherited breast cancer
非常感谢！嘿嘿，以前未接触过，刚才又去查了一下PCR-ELISA，终于有点头绪；多谢前辈！
不好意思啊，挠头；还待求助前辈，您有那两篇文章的全文吗？刚才搜了一下，没找到...
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